Fig. 1.

Ectopic overexpression of FOXO1 increases proinflammatory gene expression in TNF-α-treated HepG2 cells. HepG2 cells in six-well plates were transfected with a plasmid expressing constitutively active FOXO1-AAA or a control vector pcDNA3.1 at a fixed dose (0.4 μg/1ml Optimem). Cells were harvested for mRNA and protein analyses 24 h after transfection. A, FOXO1 mRNA levels determined by RT-PCR using β-actin as an internal control. B, FOXO1 protein levels determined by Western blotting. C, Effects of FOXO1 on proinflammatory cytokines gene expression. HepG2 cells were transfected as described above. Cells were treated 6 h after transfection with 10 ng/ml recombinant human TNF-α for 24 h, and then harvested for RT-PCR analysis of a panel of proinflammatory genes. Each of the experiments was repeated at least three times, and similar results were obtained.